Highly inducible synthesis of heterologous proteins in epithelial cells carrying a glucocorticoid-responsive vector

Hirt, Robert P.; Poulain‐Godefroy, Odile; Billotte, Jérôme; Kraehenbuhl, Jean–Pierre; Fasel, Nicolás

doi:10.1016/0378-1119(92)90687-k

Cited by 59 publications

(42 citation statements)

References 23 publications

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“…Transwells were used at days 8 to 11. Before starting a transwell assay, the tightness of the cell layer was confirmed in at least two wells as published before (20).…”

Section: Methodsmentioning

confidence: 99%

Characterization of Human Class-Switched Polymeric (Immunoglobulin M [IgM] and IgA) Anti-Human Immunodeficiency Virus Type 1 Antibodies 2F5 and 2G12

Wolbank¹,

Kunert²,

Stiegler³

et al. 2003

J Virol

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We have previously generated human monoclonal anti-human immunodeficiency virus type 1 (anti-HIV-1) antibodies 2F5IgG and 2G12IgG with an exceptional cross-clade neutralizing potential. 2F5IgG and 2G12IgG passively administrated to macaques were able to confer complete protection from both intravenous and mucosal challenge with pathogenic HIV-simian immunodeficiency virus chimeric strains and have shown beneficial effects in a phase-1 clinical trial. We now class-switched 2F5 and 2G12 to the immunoglobulin M (IgM) or IgA isotype, to enforce features like avidity, complement activation, or the potential to neutralize mucosal transmission. For this purpose we expressed functional polymeric 2F5 and 2G12 antibodies in CHO cells and evaluated their anti-HIV-1 activity in vitro. The class switch had a strong impact on the protective potential of 2F5 and 2G12. 2G12IgM inhibited HIV-1 infection of peripheral blood mononuclear cell cultures up to 28-fold-more efficiently than the corresponding IgG and neutralized all of the primary isolates tested. The 2F5 and 2G12 antibodies of all isotypes were able to interact with active human serum to inhibit viral infection. Furthermore, we demonstrated that polymeric 2F5 and 2G12 antibodies but not the corresponding IgGs could interfere with HIV-1 entry across a mucosal epithelial layer in vitro. Although polymeric 2F5 antibodies had only limited potential in the standard neutralization assay, the results from the mucosal assay suggest that 2F5 and 2G12 antibodies may have a high potential to prevent natural HIV-1 transmission in vivo.

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“…Transwells were used at days 8 to 11. Before starting a transwell assay, the tightness of the cell layer was confirmed in at least two wells as published before (20).…”

Section: Methodsmentioning

confidence: 99%

Characterization of Human Class-Switched Polymeric (Immunoglobulin M [IgM] and IgA) Anti-Human Immunodeficiency Virus Type 1 Antibodies 2F5 and 2G12

Wolbank¹,

Kunert²,

Stiegler³

et al. 2003

J Virol

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“…The sIgPKD193 construct (36) was created by fusion of a membrane expression cassette with the COOH-terminal cytoplasmic domain of murine polycystin-1 (provided by Drs. G. Waltz and G. Germino) and ligated into the glucocorticoidresponsive expression vector pLKNeo (18). Control lines were generated by stable transfections with the isolated membrane expression cassette (sIg) or vector alone.…”

Section: Methodsmentioning

confidence: 99%

The isolated polycystin-1 cytoplasmic COOH terminus prolongs ATP-stimulated Cl^–conductance through increased Ca²⁺entry

Wildman¹,

Hooper²,

Turner³

et al. 2003

American Journal of Physiology-Renal Physiology

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“…The modified ER-LBD was mutated such that it binds 4-hydroxytamoxifen (4OHT) but not 17␤-estradiol (Feil et al, 1997). To generate an expression vector for our studies with the L cells, the Moloney murine sarcoma virus (MSV) long terminal repeat (LTR) promoter was removed from the pEMSVscribe␣2 expression vector (gift of Andres Lassar, Harvard University) (Davis et al, 1987) and moved to the backbone of the pLKneo expression vector (Hirt et al, 1992), replacing the MMTV promoter present in pLKneo, but retaining the SV40-derived poly A site, plus the neomycin resistance gene for selection. This expression vector also contained the N-cadherin/ER-LBD (NcadER).…”

Section: N-cadherin/er-lbd Constructmentioning

confidence: 99%

Modulating the strength of cadherin adhesion: evidence for a novel adhesion complex

Kim

Sauer

Testa

et al. 2005

Journal of Cell Science

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Adherens junctions and desmosomes are critical for embryogenesis and the integrity of adult tissues. To form these junctions, classical cadherins interact via α- and β-catenin with the actin cytoskeleton, whereas desmosomal cadherins interact with the intermediate filament system. Here, we used a hormone-activated mutant N-cadherin expressed in fibroblasts to show the existence of a novel classical cadherin adhesion system. N-cadherin was fused at its C-terminus to a modified estrogen receptor ligand-binding domain (NcadER) that binds 4-hydroxytamoxifen (4OHT) and expressed in L cells, which lack an endogenous cadherin. Cells with the mutant cadherin (LNER cells) aggregated in the absence of 4OHT, but only in its presence formed tightly compacted aggregates like those formed by L cells expressing wild-type N-cadherin (LN cells). Compaction of LNER cells treated with 4OHT was accompanied by elevated levels of p120ctn in NcadER immunoprecipitates, compared to immunoprecipitates of non-treated cells, but without changes in α- and β-catenin, or actin. Compaction induced by 4OHT was also accompanied by increased interaction of the NcadER with the cytoskeleton and increased vimentin organization. Vimentin co-immunoprecipitated with the NcadER/catenin complex, suggesting an interaction between cadherin and vimentin. The mechanism by which vimentin interacts with the cadherin appears to involve p120ctn as it co-immunoprecipitates and colocalizes with vimentin in the parent L cells, which lack a cadherin and α- and β-catenins. Disrupting the actin cytoskeleton with cytochalasin B inhibited aggregation, whereas knocking down vimentin with specific siRNAs inhibited compaction. Based on our results we propose that a vimentin-based classical cadherin complex functions together with the actin-based complex to promote strong cell-cell adhesion in fibroblasts.

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Highly inducible synthesis of heterologous proteins in epithelial cells carrying a glucocorticoid-responsive vector

Cited by 59 publications

References 23 publications

Characterization of Human Class-Switched Polymeric (Immunoglobulin M [IgM] and IgA) Anti-Human Immunodeficiency Virus Type 1 Antibodies 2F5 and 2G12

Characterization of Human Class-Switched Polymeric (Immunoglobulin M [IgM] and IgA) Anti-Human Immunodeficiency Virus Type 1 Antibodies 2F5 and 2G12

The isolated polycystin-1 cytoplasmic COOH terminus prolongs ATP-stimulated Cl^–conductance through increased Ca²⁺entry

Modulating the strength of cadherin adhesion: evidence for a novel adhesion complex

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Highly inducible synthesis of heterologous proteins in epithelial cells carrying a glucocorticoid-responsive vector

Cited by 59 publications

References 23 publications

Characterization of Human Class-Switched Polymeric (Immunoglobulin M [IgM] and IgA) Anti-Human Immunodeficiency Virus Type 1 Antibodies 2F5 and 2G12

Characterization of Human Class-Switched Polymeric (Immunoglobulin M [IgM] and IgA) Anti-Human Immunodeficiency Virus Type 1 Antibodies 2F5 and 2G12

The isolated polycystin-1 cytoplasmic COOH terminus prolongs ATP-stimulated Cl–conductance through increased Ca2+entry

Modulating the strength of cadherin adhesion: evidence for a novel adhesion complex

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The isolated polycystin-1 cytoplasmic COOH terminus prolongs ATP-stimulated Cl^–conductance through increased Ca²⁺entry